This research will provide the scientific and engineering information upon which can be based the confident engineering design of automated laboratory analyses. Broadly applicable principles are sought as well as the specifics for particular analyses. Particular attention is being paid to cancer detection, the mucopolysaccharidoses and other heritable storage disorders, and diabetes mellitus. Identification, quantitation, and function of intracellular macromolecules associated with these diseases is being pursued by means of Biophysical Cytochemistry which employs vital, fluorescent molecular probes (e.g., acridine orange, AO) and absolute fluorescence and fluorescence-excitation microspectrophotometry, fluorescence fading, and microspectropolarimetry, as well as thermodynamic binding studies. Investigations are conducted on single unfixed cells and in vitro model systems. A fluorescence standards system has been conceived and will be developed. Traceability to the NBS will be provided. Disease-associated differences among cells and among urine samples have already been found. Possibilities for automated analyses are evident and will be implemented. Strategies for automating the detection of uterine cancer will be tested. Applications to other types of cancer will be investigated. Intracellular mucopolysaccharides (AMPS) have been identified by fluorescence fading. Applications to automated analyses will be forthcoming. Fluorescence fading provides basic chemical knowledge and promises rapid selective urine analyses for AMPS and similar molecules. Polyene antibiotics, fluorescent probes for sterols, will be used in conjunction with AO staining to study cholesterol and membrane physiology with emphasis on diabetes mellitus. Functional differences useful for automation will be sought in chronic studies of cells in culture and in perfusion chambers. Methods will be sought for quantitating the amount of an intracellular biopolymer in situ. BIBLIOGRAPHIC REFERENCES: Golden, J.F., West, S.S., Echols, C.K., and Shingleton, H.M. (1976), "Quantitative Fluorescence Spectrophotometry of Acridine Orangestained Unfixed Cells: Potential for Automated Detection of Human Uterine Cancer," J. Histochem. Cytochem. 24: 315-321.